We plan to sequence the genomes of a group of Gram-negative bacterial pathogens that are related to E. coli, making use of the completed E. coli K-12 sequence to accelerate assembly and analysis of the new genomes. The pathogens we have selected include members of E. Coli human diarrheagenic and extraintestinal strains (enteropathogenic, enterotoxigenic, enteroaggregative, uropathogenic E. coli, and K1, cause of neoatal sepsis and meningitis). The diarrheagenic strains cause diseases of major importance strains cause diseases of major importance in developing, countries, whereas the extraintestinals are frequently responsible for nosocomial or community-acquired infections in North America and Europe. In addition to these E. coli genomes we will determine the genomic sequence of Yersinia pestis KIM, causative agent of plague, Shigella flexneri 2 a, the principle agent of dysentary, and Salmonella typhi Ty2, causing Typhoid fever. These diseases are, again, of most concern in developing countries, through reservoirs do exist in other parts, including the US. Other strains of Yersinia and Shigella will be added if time permits. In a separately funded project, we have almost completed sequencing the genome of enterohemorrhagic E. coli O157:H7 EDL933, the cause of recent outbreaks of acute diarrhea and hemolytic uremic syndrome. This genome has been assembled on E. coli K-12 as a backbone, and will serve as a model for the genomics approach. Since many virulence determinants, and indeed, many "backbone" genes, are shared among the pathogens, we expect this multigenomic approach to allow rapid identification of a "pathosphere" of the virulence genes that make up the pathogenic potential of this group of bacteria. The genome center at the University of Wisconsin was established to sequence the genome of Escherichia coli K-12 strain MG1655, which has served for decades as a model organism for basic studies of biochemistry, physiology, genetics and biotechnology. The entire sequence was completed and released in GenBank in January 1997, and published in Science in September, 1997. With this sequence complete, we now plan to make use of our expertise by addressing at least eight human enteric pathogens.